EP0120694A2
Expired / Lapsed
EP
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Processes for the production of multichain polypeptides or proteins
Assignee: Celltech R&D Ltd
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Filing: 1984-03-23
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Granted: —
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Expires: 2004-03-23
Abstract
Abstract This invention relates to multichain polypeptides or proteins and processes fortheir production in cells of host organisms which have been transformed by recombinant DNA techniques. According to a first aspect of the present invention, there is provided a process for producing a heterologous multichain polypeptide or protein in a single host cell, which comprises transforming the host cell with DNA sequences coding for each of the polypeptide chains and expressing said polypeptide chains in said transformed host cell. According to another aspect of the present invention there is provided as a product of recombinant DNA technology an Ig heavy or light chain or fragment thereof having an intact variable domain. The invention also provides a process for increasing the level of protein expression in a transformed host cell and vectors and transformed host cells for use in the processes.
Inventors
- 👤 Michael Alan Boss
- 👤 John Henry Kenten
- 👤 John Spencer Emtage
- 👤 Clive Ross Wood
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 18:58
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/46; C07K16/461; C07K16/462; A; A61; A61P; A61P7/00; B; B01; B01D; B01D15/00; B01D15/08; B01D15/26; B01D15/38; B01D15/3804; B01D15/3809; C12; C12N; C12N15/00; C12N15/09; C12N15/63; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; A61K; A61K38/00; C07K2317/00; C07K2317/20; C07K2317/24
Claims Analysis
Total Claims: 38
Claim 1
Independent Claim
42 words
A process for producing a heterologous multi chain polypeptide or protein in a single host cell, which comprises transforming the host cell with DNA sequences coding for each of the polypeptide chains and expressing said polypeptide chains in said transformed host cells.
Claim 2
Dependent Claim
42 words
A process for producing a heterologous multi chain polypeptide or protein in a single host cell, which comprises transforming the host cell with DNA sequences coding for each of the polypeptide chains and expressing said polypeptide chains in said transformed host cells.
Claim 3
Dependent Claim
28 words
A process according to claim 1, wherein each DNA sequence is inserted individually into a vector and the individual vectors are used to transform the single host cell.
Claim 4
Dependent Claim
31 words
A process according to claim 1, wherein at least two of the DNA sequences are inserted into separate parts of a vector which is used to transform the single host cell.
Claim 5
Dependent Claim
17 words
A process according to claim 2 or claim 3 wherein the or each vector is a plasmid.
Claim 6
Dependent Claim
18 words
A process according to claim 4, wherein the plasmid is col El, pcRl, pBR322, RP4 or phage DNA.
Claim 7
Dependent Claim
21 words
A process according to any one of claims 1 to 5 wherein the host cell is a bacterium or a yeast.
Claim 8
Dependent Claim
16 words
A process according to claim 6, wherein the host cell is E.Coli, B. subtilis or S.cerevisiae.
Claim 9
Dependent Claim
22 words
A process according to claim 7, wherein the host cell is E.Coli strain HB101,_X1776, X2887, PS410, NRC 1 RV308, B or El1035.
Claim 10
Dependent Claim
23 words
A process according to any one of claims. 1. to 8, wherein the polypeptide chains are expressed and secreted by the host cell.
Claim 11
Dependent Claim
34 words
A process according to any one.of claims 1 to 8, wherein the polypeptide chains expressed by the host cell are solubilised under conditions which facilitate assembly of the active multi chain polypeptide or protein.
Claim 12
Dependent Claim
45 words
A process according to any one of claims I to 10, for producing an Ig molecule or an immunologically functional Ig fragment, wherein the host cell is transformed with DNA sequences coding for at least the variable domains of the Ig heavy and light chains.
Claim 13
Dependent Claim
19 words
A process according to claim 11, wherein the DNA sequences code for the complete Ig heavy and light chainsr
Claim 14
Dependent Claim
41 words
A process according to claim 11 or claim 12, for producing an Ig molecule or fragment having at least one constant domain, wherein the constant domain is derived from the same source as the variable domain to which it is attached.
Claim 15
Dependent Claim
46 words
A process according to claim 11 or claim 12, for producing an Ig molecule or fragment having at least one constant domain, wherein the constant domain is derived from a different source for that from which the variable domain to which it is attached is derived.
Claim 16
Dependent Claim
32 words
A process according to any one of claims 11 to 14, wherein the DNA coding sequences for the Ig molecule or fragment are derived from one or more monoclonal antibody producing hybridomas.
Claim 17
Dependent Claim
37 words
A process for increasing the level of protein expression in a transformed host cell comprising altering the vector DNA sequence in the region of the initiator AT G codon to one which substantially eliminates RNA secondary structure.
Claim 18
Dependent Claim
37 words
A process for increasing the level of protein expression in a transformed host cell comprising altering the vector DNA sequence in the region of the initiator AT G codon to one which substantially eliminates RNA secondary structure.
Claim 19
Dependent Claim
24 words
A process according to claim 16 wherein the DNA sequence is altered to minimise the absolute value of the Δ G value.of the sequence.
Claim 20
Dependent Claim
17 words
A heterologous multi chain polypeptide or protein produced by recombinant DNA technology from a single host cell.
Claim 21
Dependent Claim
17 words
A heterologous multi chain polypeptide or protein produced by recombinant DNA technology from a single host cell.
Claim 22
Dependent Claim
19 words
An Ig molecule or an immunologically functional Ig fragment produced by recombinant DNA technology from a single host cell.
Claim 23
Dependent Claim
19 words
An Ig molecule or an immunologically functional Ig fragment produced by recombinant DNA technology from a single host cell.
Claim 24
Dependent Claim
29 words
An Ig molecule or fragment according to claim 19, including at least one constant domain derived from the same source as the variable domain to which it is attached.
Claim 25
Dependent Claim
35 words
An Ig molecule or fragment according to claim 19, including at least one constant domain derived from a source different from the source from which the variable domain to which it is attached was derived.
Claim 26
Dependent Claim
20 words
An Ig fragment according to claim 20 or claim 21, comprising only one constant domain attached to each variable domain.
Claim 27
Dependent Claim
25 words
An Ig molecule according to claim 20 or claim 21, comprising sufficient constant domains attached to each variable domain to produce a complete Ig molecule.
Claim 28
Dependent Claim
22 words
An Ig heavy or light chain or a fragment thereof having a complete variable domain, as a product of recombinant DNA technology.
Claim 29
Dependent Claim
22 words
An Ig heavy or light chain or a fragment thereof having a complete variable domain, as a product of recombinant DNA technology.
Claim 30
Dependent Claim
16 words
A vector for use in a process according to any one of claims 1 to 17.
Claim 31
Dependent Claim
26 words
A vector for use in transforming a host cell to enable it to produce a molecule according to any one of claims 18 to 24 -
Claim 32
Dependent Claim
13 words
A vector according to claim 25 or claim 26, which is a plasmid
Claim 33
Dependent Claim
16 words
A host cell transformed with a vector according to any one of claims 25 to 27.
Claim 34
Dependent Claim
20 words
A therapeutic composition comprising a molecule according to any one of claims 18 to 24 and a pharmaceutically acceptable carrier.
Claim 35
Dependent Claim
28 words
A process for preparing a composition according to claim 29, comprising mixing a molecule according to any one of claims 18 to 24 with a pharmaceutically acceptable carrier.
Claim 36
Dependent Claim
21 words
An affinity chromatography medium comprising a molecule according to any one of claims 18 to 24 immobilised on a chromatographic medium.
Claim 37
Dependent Claim
29 words
An immunological assay method in which the or each antigen binding Ig molecule or fragment is a molecule or fragment according to any one of claims 19 to 24.
Claim 38
Dependent Claim
36 words
33 . An Ig molecule or fragment according to any one of claims 18 to 24 having attached thereto a cytotoxic, enzymic or structural polypeptide moiety synthesised in the cell with the Ig molecule or fragment
References Cited (Backward Citations)
No citation records indexed.
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